Download Functional subsets of lymphocytes

Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
J. clin. Path., 32, Suppl. (Roy. Coll. Path.), 13, 59-62
Functional subsets of lymphocytes
P. C. L. BEVERLEY
From the Imperial Cancer Research Fund Human Tumour Immunology Group, University College
Hospital Medical School, London
In this review I shall survey various properties of
lymphocytes which allow clear distinctions to be
drawn between different lymphocyte subsets. I shall
discuss only thymus-derived (T) lymphocytes since
these cells appear to play a key role in regulating the
activities of other lymphoid cells (B cells and macrophages) as well as carrying out effector functions
themselves. In the first section I shall discuss murine
T lymphocytes since most of the most decisive
experimental studies have been carried out in this
species, while the concluding section of the review
will deal with human T lymphocyte heterogeneity.
Thymus
TIl
Thy-I
Ly-123+
Bone marrow
T stem cell
Peripheral T cells
Thy-I
TLThy- I
Ly- I
TL-
Ly-I+3OI/. Ly-123+50°/. Ly-23+ 1OI/.
Ly
Fig. 1 Differentiation of T cells.
T Lymphocyte heterogeneity in the mouse
described (Beverley, 1977; Simpson and Beverley,
1977). However, it illustrates the important point
that during differentiation of T cells from stem cells
resident in the bone-marrow to mature peripheral T
cells there is a major reorganisation of the cell surface. Some markers (for example, TI) are lost while
others appear (for example, Ala-1 which is present
only on mature effector cells of both T and B
lineages). In the peripheral lymphoid tissues the
Ly-1, 2, and 3 markers allow a division of the T cells
into three major subsets with different surface
expression of the three antigens (phenotype) and
distinct functional properties (Fig. 1).
Although the realisation that different subsets of
lymphocytes served different functions-for example,
Table 1 Murine T lymphocyte surface antigens
that B cells produce antibody and T cells mediate
delayed hypersensitivity-preceded theuse of antisera
Reference
Linkage group
Antigen
to differentiation antigens, use of these reagents has
IX
allowed an analysis of T cell function which was
H-2
Boyse and Old, 1968
IX
la
Frelinger et al., 1974
previously impossible. Particularly enlightening are
IX
TL
Boyse and Old, 1969
the studies by Cantor and Boyse (1975a, b) of mixed
Raff and Wortis, 1970
II
Thy-1
XII
Boyse et al., 1968
Ly-1
culture (MLC) responses. Their findings
lymphocyte
XI
Boyse et al., 1968
Ly-2
may be summarised as follows.
XI
Boyse et al., 1971
Ly-3
?
Ala-1
Feeney and Hammerling, 1976
(1) The major portion of the proliferative
response in MLC is due to Ly-1+ cells.
(2) The Ly-1+ cells respond to alloantigens coded
Using a panel of antisera allows the construction in the I region of the major histocompatibility
of a scheme for T cell differentiation such as that complex (MHC).
(3) Cytotoxic effector cells generated in MLC and
given in Fig. 1. This is a considerable over-simplification since many other T cell markers have been their precursors are Ly-23+.
59
The most useful tool for separation of T cells into
different subsets has been the use of antisera to cell
surface differentiation markers. Cells bearing a
particular antigen may be killed by antiserum and
complement or they may be detected and separated
by such techniques as indirect immunofluorescence.
Although T lymphocyte differentiation antigens may
be detected by both heteroantisera and alloantisera
(reviewed by Beverley, 1977) the most useful markers
have been those detected by alloantisera. Table 1
lists some of the markers that have been particularly
useful in elucidating the pathway of T cell differentiation in the mouse.
Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
60
P. C. L. Beverley
Thymus
Ly- 123
Ly-1v *
Helpers
Amplifiers
I restricted
Ly-23+
Suppressors
Killers
KID restricted
Fig. 3 T cell differentiation.
lymphocytes recognise extrinsic antigen in association with self MHC antigens and, furthermore, that
there are two major subtypes of T cells (Fig. 3). The
Ly-1+ cells are helper/amplifiers for other cell types
and respond to I region antigens, while Ly-(1)23+
cells are suppressor/killers and K/D responsive. In
addition, Fig. 3 emphasises the interactions between
these two broad categories of T cell.
Fig. 2 The mixed lymphocyte culture (MLC) responses.
Human lymphocyte subsets
(4) The target antigens of the cytotoxic cells are
H-2K and D.
(5) Generation of an optimal cytotoxic response
requires the presence of Ly-1+ cells.
These important findings are summarised diagrammatically in Fig. 2. The study established not only
that different cells mediate different functions but
also that the different cells respond to antigens coded
in different regions of the MHC. In addition the
importance of cell-cell interactions in generation of
effector cells was emphasised.
Now it could be argued that the MLC is a nonphysiological in-vitro culture artefact, and one might
ask what relevance MHC recognition has to host
defence against bacteria and viruses? The now
classical experiments of Zinkernagel and Doherty
(reviewed in Doherty et al., 1976) provide an answer.
They show that cytotoxic T cells, immune to viral
antigens, recognise not just the viral antigens but
viral antigens in association with self MHC antigens.
The relevant MHC antigens are coded in the K and
D regions. In addition it has now been shown that
anti-viral cytotoxic cells are Ly-23+ (Pang et al.,
1976).
Does this self + antigen recognition apply only
to Ly-23+ killer cells? Somewhat more indirect
evidence than that for killer cells shows that helper
cells for antibody production recognise soluble
protein antigens in association with I region coded
antigens (Erb et al., 1976). The helper cells have also
been shown to have the Ly-1+ phenotype (Feldmann
et,£., 1975). In other experiments it has also been
demonstrated that the Ly-1+ cells which mediate
delayed type hypersensitivity reactions are also I
region restricted (Miller et al., 1976).
Taken together these data suggest that all T
In this section I briefly review our present understanding of human lymphocyte heterogeneity. Fig. 4
summarises the distribution of cell phenotypes that
may be found in human peripheral blood as distinguished by a number of widely used markers.
These separation techniques have allowed considerable advances in understanding of human cellular
immune responses but do not allow adequate
definition of the non-T (non-E-rosetting) subsets nor
subfractionation of the T cells.
O
10
20
30 40
50 60
80 90 100
70
E
SMI 19l
_c~_
E
//
/
19No
15
375
T
MONO
10
250
B
EAkc
/0
Cells/mrrn3
75
Fig. 4 Normal lymphocyte phenotypes.
Three sets of markers have allowed a start oi.
investigations of human T cell subsets. The first is
the use of IgM and IgG rosetting. Two subpopulations of T cells can be identified by the use of ox red
blood cells coated with IgM or IgG antibody. The
T.M cells comprise about 55-60% and the T.G cells
10% of peripheral blood T cells (Moretta et al.,
1976). Complete functional studies of these subsets
have not so far been reported but they show
Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
Functional subsets of lymphocytes
61
Table 2 Summary of data derivedfrom studies with
juvenile rheumatoid arthritis (JRA) sera and anti-THJ
and TH2 sera
40*
Antigens
Function/response
MLC
HELP
CML
Suppression
Soluble antigen
20-
10Control
----------
TH1+
+
+
ND
-
TH2+
_
+
+
-
JRA+
_
ND
+
+
---
not fall clearly into either category. Similarly in the
mouse it is unclear which subset of T cells is most
important in this response. In summary, it appears
that distinctions, both functional and phenotypic,
Fig. 5 Cytotoxicity ofjuvenile rheumatoid arthritis
between the major lymphocyte subsets have been
(JRA) sera. Three JRA sera were assayed on nylon
conserved during mammalian evolution.
column purified human PBL (> 90 %E+) using a
Since it is now clear that major subsets of human
two-stage s Cr release assay with absorbed rabbit serum
lymphocytes having distinct surface and functional
as a source of complement.
properties can be defined, we might ask the relevance
of these to the pathogenesis of disease. As yet data
differences in response to PHA. More importantly, it are scanty, but there is already evidence for dishas also been demonstrated that the T cells required turbances in numbers of T. M and T. G cells in certain
to potentiate a B cell response to pokeweed mitogen immunodeficiency and autoimmune disorders
reside in the T.M subset. T. G cells, on the other hand, (Moretta et al., 1977) and in multiple sclerosis
(Santoli et al., 1978). Preliminary evidence also
inhibit this response (Moretta et al., 1976).
There are more complete data on the use of two suggests that JRA patients with detectable autodifferent types of antisera to distinguish T cell sub- antibody lack circulating suppressor cells (Schlosssets. Schlossman and his colleagues have investigated man et al., 1978).
Rapid progress may be expected in this area when
the properties of sera from patients suffering from
juvenile rheumatoid arthritis (JRA). Active sera have the existing markers are improved. Present heteroan autoantitody capable of killing up to 40% of sera suffer from limited availability, difficulty in
human peripheral blood T cells (Strelkauskas et al., reproducing successful immunisations, and multiple
1978). In our experience the sera are rather more specificities present in the same serum. JRA sera are
heterogeneous (Fig. 5) giving variable levels of not readily available and only untreated patients
plateau killing of peripheral blood T lymphocytes. have usable titres. Furthermore, the number of cells
Nevertheless, in Schlossman's hands several high killed may vary (Fig. 5), suggesting multiple specikilling sera have given repeatable results in studies ficities within the sera. Perhaps the most promising
of lymphocyte function (Strelkauskas et al., 1978; avenue for advance is the use of somatic cell
hybridisation to produce monoclonal antisera
Schlossman et al., 1978).
An alternative approach is to develop specific (Kohler and Milstein, 1976). Already an antigen
heteroantisera. Rabbit antisera to T lymphocytes present on human thymocytes but not peripheral T
may be absorbed with autologous B lympho- cells has been defined (McMichael et al., 1978)
cytes (Evans et al., 1977), or T leukaemia cells (Evans which is perhaps equivalent to the mouse TL antigen.
et al., 1978) to derive sera recognising subsets of T It is to be expected that peripheral T and B cell
lymphocytes. In this way two markers TH1 and markers will shortly be described.
TH2 have been identified. Table 2 summarises (and
simplifies) the data derived from studies with JRA
sera and anti THI and TH2 sera. It appears that References
most of the functions measured can be ascribed to
P. C. L. (1977). Lymphocyte heterogeneity.
two major subsets of lymphocytes-the TH1+ Beverley,
In
B
T Cells in Immune Recognition, edited by F.
and
helper/amplifier, which appears broadly equivalent
Loor and G. E. Roelants, pp. 35-57. Wiley, London.
to the mouse Ly-1+ cell, and the TH2+ JRA+ sup- Boyse, E. A., Itakura, K., Stockert, E., Iritani, C. A., and
pressor/killer, broadly equivalent to the mouse
Miura, M. (1971). Ly-C: a third locus specifying
Ly-23+. It is interesting that the cells which give an
alloantigens expressed only on thymocytes and
in-vitro proliferative response to soluble antigen do
lymphocytes. Transplantation, 11, 351-353.
0
1:16
Serum dilution
1:4
1:64
1:256
1:1024
Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
62
P. C. L.
Beverly
Boyse, E. A., Miyazawa, M., Aoki, T., and Old, L. J. Kohler, G., and Milstein, C. (1976). Derivation of
(1968). Ly-A and Ly-B: two systems of lymphocyte
specific antibody-producing tissue culture and tumor
isoantigens in the mouse. Proceedings of The Royal
lines by cell fusion. European Journal of Immunology, 6,
Society Series B, 170, 175-193.
511-519.
Boyse, E. A., and Old, L. J. (1969). Some aspects of McMichael, A. J., Pilch, J. R., Galfre, G., Mason, D. Y.,
normal and abnormal cell surface genetics. Annual
Fabre, J. W., and Milstein, C. (1979). A human
Review of Genetics, 3, 269-290.
thymocyte antigen defined by a hybrid myeloma
Cantor, H., and Boyse, E. A. (1975a). Functional
monoclonal antibody. European Journal of Immunology,
subclasses of T lymphocytes bearing different Ly
9,205-210.
antigens. 1. The generation of functionally distinct T Miller, J. F. A. P., Vadas, M. A., Whitelaw, A., and
cell subclasses is a differentiative process independent
Gamble, J. (1976). Role of major histocompatibility
of antigen. Journal of Experimental Medicine, 141,
complex gene products in delayed type hypersensitivity.
1376-1389.
Proceedings of the National Academy of Sciences, 73,
Cantor, H., and Boyse, E. A. (1975b). Functional sub2486-2490.
classes of T lymphocytes bearing different Ly antigens. Moretta, L., Ferrarini, M., Mingari, M. C., Moretta, A.,
II. Co-operation between subclasses of Ly cells in the
and Webb, S. R. (1976). Subpopulations of human T
generation of killer activity. Journal of Experimental
cells identified by receptors for immunoglobulins and
Medicine, 141, 1390-1405.
mitogen responsiveness. Journal of Immunology, 117,
Doherty, P. C., Blanden, R. V., and Zinkernagel, R. M.
2171-2174.
(1976). Specificity of virus-immune effector T cells for Moretta, L., Mingari, M. C., Webb, S. R., Pearl, E. R.,
H-2K or H-2D compatible interactions: implications
Lydyard, P. M., Grossi, C. E., Lawton, A. R., and
for H-antigen diversity. Transplantation Reviews,
Cooper, M. D. (1977). Imbalances in T cell sub29, 89-124.
populations associated with immunodeficiency and
Erb, P., Feldmann, M., and Hogg, N. M. (1976). Role
autoimmune syndromes. European Journal of Imof macrophages in the generation of T helper cells.
munology, 7, 595-700.
IV Nature of genetically related factor derived from Moretta, L., Webb, S. R., Grossi, C. E., Lydyard, P. M.,
macrophages incubated with soluble antigens. European
and Cooper, M. D. (1977). Functional analysis of two
Journal of Immunology, 6, 365-372.
human T-cell subpopulations. Help and suppression of
Evans, R. L., Beard, J. M., Lazarus, H., Schlossman,
B-cell responses by T cells bearing receptors for IgM
S. F., and Chess, L. (1977). Detection, isolation, and
or IgG. Journal of Experimental Medicine, 146, 184-200.
functional characterization of two human T-cell Pang, T., McKenzie, I. F. C., and Blanden, R. V. (1976).
subclasses bearing unique differentiation antigens.
Co-operation between mouse T-cell subpopulations in
Journal of Experimental Medicine, 135, 221-233.
the cell-mediated response to a natural poxvirus
Evans, R. L., Lazarus, H., Penta, A. C., and Schlossman,
pathogen. Cellular Immunology, 26, 153-159.
S. F. (1978). Two functionally distinct subpopulations Raff, M. C., and Wortis, H. H. (1970). Thymus
of human T cells that collaborate in the generation of
dependence of 0-bearing cells in the peripheral lymphoid
cytotoxic cells responsible for cell-mediated
tissues of mice. Immunology, 18, 931-943.
lympholysis. Journal of Immunology, 120, 1423-1428.
Santoli, D., Moretta, L., Lisak, R., Gilden, D., and
Feeney, A. J., and Hammerling, U. (1976). Differentiation
Koprowski, H. (1978). Imbalances in T cell subof murine T and B lymphocytes from resting to effector
populations in multiple sclerosis patients. Journal of
cells, as characterised by ALA-1 (activated lymphocyte
Immunology. 120,1369-1371.
antigen-i). In Proceedings of the Second International Schlossman, S. F., Evans, R. L., and Strelkauskas, A. J.
Congress on Cell Differentiation. North Holland
(1978). Human T-cell subsets with regulatory functions.
Publishing, Amsterdam.
In Inserm Symposium 8, edited by B. Serrou and C.
Feldmann, M., Beverley, P. C. L., Dunkley, M., and
Rosenfeld, North-Holland, Amsterdam.
Kontiainen, S. (1975). Different Ly antigen phenotypes Simpson, E., and Beverley, P. C. L. (1977). T cell
of in-vitro induced helper and suppressor cells. Nature
subpopulations. Progress in Immunology, 3, 206-216.
(London), 258, 614-616.
Strelkauskas, A. J., Schauf, V., Wilson, B. S., Chess, L.,
Frelinger, J. A., Niederhuber, J. E., David, C. S., and
and Schlossman, S. F. (1978). Isolation and characterSchreffiler, D. C. (1974). Evidence for the expression of
ization of naturally occurring subclasses of human
la (H-2 associated) antigens on thymus derived
peripheral blood T cells with regulatory functions.
lymphocytes. Journal of Experimental Medicine, 140,
Journal of Immunology, 120, 1278-1282.
1273-1284.
Downloaded from http://jcp.bmj.com/ on June 17, 2017 - Published by group.bmj.com
Functional subsets of
lymphocytes.
P C Beverley
J Clin Pathol 1979 s3-13: 59-62
doi: 10.1136/jcp.s3-13.1.59
Updated information and services can be found at:
http://jcp.bmj.com/content/s3-13/1/59.citation
These include:
Email alerting
service
Receive free email alerts when new articles cite this
article. Sign up in the box at the top right corner of
the online article.
Notes
To request permissions go to:
http://group.bmj.com/group/rights-licensing/permissions
To order reprints go to:
http://journals.bmj.com/cgi/reprintform
To subscribe to BMJ go to:
http://group.bmj.com/subscribe/